谷氨酰胺缓解中华鳖免疫应激反应作用机理的研究
|
摘要
中华鳖(Trionyx sinensis)是一种古老的次生性水生爬行动物,具有重要的研究价值和经济价值。80年代以来,不断扩大的市场需求刺激了养鳖业的迅猛发展。各种环境因素、人为的管理因素以及病原微生物的影响,使中华鳖经常处于应激状态,致使其抗病能力减弱,肌肉品质下降。Gln是一种条件性必需氨基酸,具有重要的营养和药理作用。本论文通过腹腔注射E.coli LPS初步建立中华鳖免疫应激模型,并通过注射或在饲料中添加Gln,从机体内分泌、抗氧化能力、免疫功能和生产性能等方面考察Gln对中华鳖的免疫保护作用及其机理。为深入探索免疫应激机理,促进中华鳖的健康养殖提供理论与实践依据。本论文分三章,设计并实施了以下7个试验。
第一章,应用LPS诱导中华鳖免疫应激的研究。分设三个试验,即试验一至试验三。试验一,选择体重接近的健康中华鳖42只(339.46±25.54)g,随机分成7组(A组-G组),每组设2个重复,每个重复3只。上午10:00时,分别给A组-G组鳖从腹腔注射LPS(剂量分别为0,50,100,250,500,1000,5000μg/kg BW),LPS用0.5mL 0.7%NaCl配制成注射液。下午3:00时,将各组鳖断头取血,用0.5%肝素钠抗凝,2000r/min离心10min制备血浆。用放射免疫法测定血浆COR含量。结果表明:E组(500μg/kg BW)血浆COR水平极显著高于A组(对照组)、B组(50μg/kg BW)和C组(100μg/kg BW)(P＜0.01),显著高于D组(250μg/kg BW)(P＜0.05)。试验二,选择中华鳖30只(329.66±23.98)g,随机分成5组(A组-E组),每组设2个重复,每个重复3只。上午10:00时,A组(对照组)注射0.7%NaCl,B组-E组(试验组)分别注射500μg LPS/kg BW,注射剂量均为0.5mL。分别于注射当日下午3:00时A组、B组),注射后1 d(C组)、4 d(D组)和7 d(E组)的下午3:00时断头取血,制各血浆。用放射免疫法测定各血浆样品COR、TNF-α、IL-1β和IL-6的含量;用免疫浊度法测定血浆样品IgM、IgG和IgA的含量。结果表明:给中华鳖成鳖腹腔注射500μg LPS/kg BW 5h时,血浆COR、TNF-α和IL-1β的含量显著或极显著地高于对照组(P_(COR)＜0.01,P_(TNF-α)＜0.05,P_(IL-1β)＜0.01);注射1 d时,血浆IL-6的含量极显著高于对照组(P＜0.01);注射4 d时,血浆IgM的含量显著高于对照组(P＜0.05)。试验三,选择幼鳖54只(106.46±19.53)g,随机分成3组(A组-C组),每组设3个重复,每个重复6只。A组(对照组)注射0.7%NaCl,B组注射500μg LPS/kg BW,C组注射1 000μg LPS/kg BW。LPS用0.2mL 0.7%NaCl配制成注射液。试验2周后测试各组鳖的增重率和饲料系数。结果表明:对照组和500μg LPS/kg BW组幼鳖的增重率、饲料系数未出现显著差异,饲养过程中亦未观察到明显的行为变化。但1000μg LPS/kg BW组幼鳖的摄食能力减弱,增重率极显著降低(P＜0.01),且注射处理后反应强烈,相互咬噬裙边现象严重。综上,通过对不同剂量的LPS处理5 h时中华鳖成鳖血浆COR含量变化规律的观察(试验一),对注射500μg LPS/kg BW后不同时段成鳖血浆COR、TNF-a、IL-1β、IL-6、IgM、IgG和IgA含量变化的比较(试验二),以及对比注射不同剂量LPS后幼鳖摄食、生长和行为学上的变化(试验三),可见腹腔注射500μgLPS/kg BW能诱导中华鳖成鳖产生免疫应激反应;腹腔注射1000μgLPS/kg BW可显著抑制幼鳖生长;血浆COR含量的变化可作为监测中华鳖免疫应激反应的重要指标之一。
第二章,Gln缓解中华鳖免疫应激反应机制的研究。分设三个试验,即试验四至试验六。试验四,选择中华鳖60只(317.86±22.57)g,随机分为3组,A组设4个重复,每个重复3只,B、C组设8个重复,每个重复3只。上午10:00时,A组注射0.7%NaCl,B组注射500μg LPS/Kg BW,C组注射500μg LPS+100mgGln/Kg BW。LPS和Gln用0.5mL 0.7%NaCl配制成注射液。分别于注射当日(A组-C组)、注射后1 d(A组-B组)、4 d(A组-B组)和7 d(A组-C组)的下午3:00时从各组随机取2个重复6只鳖断头取血,制备血浆;解剖取相同部位肝脏组织20 g。检测不同时段(5h,1d,4d,7d)血浆COR、TNF-α、IL-1β、IL-6、GSH、GSH-PX、MDA以及肝脏PRO、GSH、GSH-PX、MDA含量的变化。结果表明:注射LPS后,血浆中COR、TNF-α、IL-1β、IL-6、MDA和肝脏组织PRO、MDA含量在相应时段显著或极显著升高(P＜0.01),而血浆GSH含量显著降低(P＜0.05);补充Gln不仅缓解了血浆COR、TNF-α、IL-1β和IL-6等的上升趋势,减缓了血浆GSH的下降,而且使肝脏MDA水平极显著降低。提示Gln可通过抑制有关细胞因子(如TNF-α、IL-1β、IL-6)和糖皮质激素(如COR)的分泌,以及增强机体的抗氧化能力而对免疫应激的中华鳖产生保护作用。试验五,选择中华鳖72只(332.35±13.75)g,随机分成4组(A组-D组),每组设6个重复,每个重复3只。A组(对照组)喂基础饲料,1周后腹腔注射0.7%NaCl;B组喂基础饲料,1周后腹腔注射500 ug LPS/kg BW;C组在基础饲料中添加2%Gln,1周后腹腔注射500 ug LPS/kgBW;D组在基础饲料中添加2%Gln,1周后腹腔注射0.7%NaCl。上午10:00时注射,注射剂量均为0.5mL。分别于注射处理后的第4d、11d和25d下午3:00从各组随机取2个重复共6只鳖称重,全身灭菌,断头取血。取表层淋巴细胞检测CD4~+/CD8~+和淋巴细胞转化率;取血浆测定IgM,IgG,lea和溶菌酶含量;解剖取小肠15cm(自幽门始),用1.5 mL0.7%NaCl反复灌洗,3000r/min离心10min,取上清液检测S-IgA。结果表明:免疫应激极显著提高中华鳖成鳖血浆IgM水平(P＜0.01),补充Gln可使LPS诱导的血浆IgM水平显著降低(P＜0.01);免疫应激抑制了中华鳖的细胞免疫功能(P＜0.05),补充Gln能缓解免疫应激对中华鳖细胞免疫功能的抑制(P＜0.05);补充Gln能显著促进肠道S-IgA的分泌(P＜0.01)。试验六,选择中华鳖36只(318.53±10.26)g,随机分成6组(A-F组),每组设2个重复,每个重复3只。A组注射0.7%NaCl;B组注射500μgLPS/kg BW;C组注射100mgGln/kgBW;D组注射100mgGln+8mgQ/kgBW;E组注射500μgLPS+100mgGln/kgBW;F组注射500μgLPS+100mgGln+8mgQ/kgBW。LPS、Gln和Q用0.5mL 0.7%NaCl配制成注射液。上午10:00时注射,下午4:00时分别取脾脏、肝脏和肠道组织,采用免疫组化法检测HSP70的表达。取血浆测定各组COR含量的变化。结果表明:注射500 ug LPS/kg BW显著促进了中华鳖肝脏和脾脏组织HSP70的合成,而注射100 mg Gln/kg BW或8 mg Q/kg BW抑制了中华鳖肠道、肝脏和脾脏组织HSP70的表达;Gln对免疫应激中华鳖的保护作用,与其诱导机体组织HSP70的表达无关。综合试验四、试验五和试验六的结果,表明Gln可通过抑制炎性细胞因子(如TNF-α、IL-1β、IL-6)和糖皮质激素(如COR)的分泌、调节免疫功能以及增强机体的抗氧化能力等途径对免疫应激中华鳖产生保护作用。
第三章,Gln/LPS对中华鳖生产性能的影响。设计一个试验,即试验七,选择中华鳖72只(101.92±17.53)g,随机分成4组(A-D组),每组设6个重复,每个重复3只。A、B组用基础饲料饲喂;C、D组饲料中添加2%Gln。第8d B、C组注射1000ugLPS/kg BW,A、D组注射0.7%NaCl。第51d进行同样处理。第95d检测各组鳖的增重率、相对生长率、饲料系数,脂体比、裙边比、肌肉的水分、粗蛋白、粗脂肪和氨基酸含量,取小肠、肝脏和脾脏切片,、观察其组织结构变化。结果表明:腹腔注射1000 ug LPS/kg BW,使幼鳖肠绒毛发育受损,肝细胞萎缩变性,显著抑制其生长,并降低肌肉品质。添加2%Gln,能促进肠道发育,显著促进生长,阻止免疫应激对中华鳖肝脏的损伤,改善免疫应激导致的中华鳖生产性能的下降。
Trionyx sinensis,an old secondary aquatic reptiles,are of research importance. Since 1980s,the increasing market demand has stimulated the rapid development of Trionyx sinensis breeding.Trionyx sinensis is always under the state of stress under the impact of various kinds of environment,human management and pathogeny,so that its ability to resist disease and muscle quality are reduced.Gln is a conditionally necessary amino acid,which has not only nutrition,but also important pharmacological action.The thesis sets up the mode of the immunological stress in Trionyx sinensis by using E.coli LPS,and makes an on-the-spot investigation into the protection and mechanism of Gln on internal secretion of the mechanism,ability to resist oxidation,immunologic function, and production performance of the immunological stressed Trionyx sinensis by injecting and adding Gln in the fodder.In order to profoundly probe into immunological stress mechanism,the application of Gln to strengthen the immunological protection lays a theoretical foundation for the healthy cultivation of Trionyx sinensis.
In the thesis,seven tests are set for the research on easing of immunological stress mechanism in Trionyx sinensis.
Forty-two healthy Trionyx sinensises(339.46±25.54)g whose weight is close to each other are chosen in test one and divided into seven groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.Each Trionyx sinensis was respectively injected into different doses of LPS from abdominal cavity,and heads were cut to get plasma after 5h. COR content in the blood plasma was measured using radioimmunoassay.It is proved that the plasma COR level content in group E is strikingly higher than that in group A(comparison group),B(50μg/kg BW) and C(100μg/kg BW)(P＜0.01),and is much higher than that in group D(250μg/kg BW)(P＜0.05).Thirty healthy Trionyx sinensises (329.66±23.98)g are chosen in test two and divided into five groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.We injected 0.7%NaCl in the comparison group and 500μgLPS/kgBW in each test group respectively,and cut their heads to get plasma at the same time after 5h、1d、4d and 7d.We measured COR、TNF-α、IL-1βand IL-6 content in each sample blood plasma using radio-immunoassay, and IgM、IgG and IgA content in the sample blood plasma using immunoturbidimetery.It is proved that when injected 500μgLPS/kgBW in the abdominal cavity of mature Trionyx sinensis for five hours,the COR、TNF-αand IL-1βcontent is much higher or strikingly higher than that in the comparison group(P_(COR)＜0.01,P_(TNF-α)＜0.05,P_(IL-1β)＜0.01),if injected for 1d,IL-6 content in the blood plasma is strikingly higher than that in the comparison group(P＜0.01),if injected for 4d,the IgM content in the blood plasma is much higher than that in the comparison group(P＜0.05).Fifty-four young Trionyx sinensises(106.46±19.53) g are chosen in test three,and divided into three groups at random,with 3 subgroups and 6 Trionyx sinensis in each subgroup.They were respectively injected into 0.7%NaCl,500μgLPS/kgBW and 1000μgLPS/kgBW.We tested weight gain ratio(WGR) and feed conversion ratio(FCR) of each group after two week's test.It is proved that there is neither obvious difference in WGR and FCR of young Trionyx sinensis in comparison group and group 500μg LPS/kg BW,nor obvious behavior difference in the process of feeding.However,young Trionyx sinensis in group1000μgLPS/kg BW has weakened food intake ability and remarkably decreased WGR(P＜0.01).Furthermore, there is strong reaction,frequent aggression of mutual biting skirt lateral after injection.In conclusion,through COR content change in the blood plasma of mature Trionyx sinensis by injecting different doses of LPS after 5h(test one),change of the COR、TNF-a、IL-1β、IL-6、IgM、IgG and IgA content in th blood plasma at different periods after injection of 500μg LPS/kg BW(test two),and changes in the food intake,growth and behavioristics of young Trionyx sinensis after injection of different doses of LPS(test three),the injection of 500μgLPS/kg BW into the abdominal cavity could cause immunological stress reaction in mature Trionyx sinensis,while injection of 1000μg LPS/kg BW into the abdominal cavity could inhibit its growth.in young Trionyx sinensises and COR content in the blood plasma could be regarded as one significant index to supervise the immunological stress reaction of Trionyx sinensis.
Sixty Trionyx sinensises(317.86±22.57) g are chosen in test four and divided into three groups at random,with 4 subgroups and 3 Trionyx sinensis in each subgroup in group A,with 8 subgroups and 3 Trionyx sinensis in each subgroup in group B and C.group A,B and C were respectively injected 0.7%NaCl、500μgLPS/Kg BW、500μg LPS+100mgGln /Kg BW into abdonamal cavity to inspect the content variation of COR、TNF-α、IL-1β、IL-6、GSH、GSH-PX、MDA in blood plasma and protein、GSH、GSH-PX、MDA in lever at different times.The result indicates that after injection of LPS,the content of COR、TNF-α、IL-1β、IL-6、MDA in blood plasma and PRO、MDA in the liver organization strikingly rise at the corresponding period(P＜0.01),while the GSH content in blood plasma remarkably lower(P＜0.05).The supplement of Gin not only eases the rising trend of COR、TNF-α、IL-1βand IL-6,the decrease of GSH in the blood plasma,but also prominently lowers the MDA level of liver.Gln could protect the immunological stressed Trionyx sinensis by restraining the secretion of concerned cell factors such as TNF-α、IL-1β、IL-6,glucocorticoid(COR) and improving the oxidatin resistance ability. Seventy-two Trionyx sinensises(332.35±13.75) g are chosen in test five and divided into four groups at random,with 6 subgroups and 3 Trionyx sinensis in each subgroup.Group A and B are fed with basal feed,group C and D are fed with basal feed plus 2%Gln.After being fed for one week,group B and C are injected into 500ugLPS/kgBW,group A and D are injected into 0.7%NaCl.4d,11d and 25d After injection the content of CD_4~+/CD_8~+ in peripheral blood,transformation ratio of T lymph,IgM、IgG、IgA in the blood plasma, muramidase and S-IgA content in intestinal canal were examined.The result proves that the immunological stress caused by LPS strikingly improves the IgM level(P＜0.01) in the blood of Trionyx sinensis.Supplement of Gln not only improves the immunologic function of cells and intestinal canal,but also prohibits the rising of IgM level in the blood plasma conducted by LPS.Thirty-six Trionyx sinensises(318.53±10.26)g are chosen in test six and divided into six groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.We used LPS、Gln and Q to treatt them.After 6h,we got spleen,liver and intestinal canal organization and inspected the expression of HSP70 by immunochemistry.At the same time,we inspected the COR content variety of each group. It is proved that injection of 500ugLPS/kgBW remarkably improves the expression of HSP70 in spleen and liver organization of Trionyx sinensis,while injection of 100mgGln/ kgBW or 8mgQ/kgBW inhibits the expression of HSP70 in intestinal canal,spleen and liver organization.To sum up,Gln could protect the immunological stressed Trionyx sinensis by regulating the immunological function,inhibiting the secretion of inflammatory cell factor and improving the oxidation resistance ability,while the protection might have nothing to do with the expression of HSP70.
Seventy-two Trionyx sinensises(101.92±17.53) g are chosen in test seven and divided into four groups at random,with 6 subgroups and 3 Trionyx sinensis in each subgroup.Group A and B are fed with basal feed,and group C and D are fed with basal feed plus 2%Gln.On the eighth day,group B and C are injected into 1000ugLPS/kg BW and 0.7%NaCl is injected into group A and D.On 51th day,they are under the same treatment,on 95th day,we inspected the weight gain ratio,comparison growth ratio, feed conversion ratio,the content of body fat ratio,body skirt ratio,muscle moisture, coarse protein,coarse fat and amino acid,and got small intestine,liver and spleen to observe their structure variation.It is proved that injection of 1000ugLPS/kgBW caused the damage of the villus growth of the intestine,the shrinkage and degeneration of liver cells,which strikingly inhibits the growth of the young Trionyx sinensis and lowers the quality.Supplement of 2%Gln could improve the growth of intestine,remarkably enhance the growth,restrain the damage of immunological stress to the liver of Trionyx sinensis,and improve the production performance of Trionyx sinensis reduceded by immunological stress.
第一章,应用LPS诱导中华鳖免疫应激的研究。分设三个试验,即试验一至试验三。试验一,选择体重接近的健康中华鳖42只(339.46±25.54)g,随机分成7组(A组-G组),每组设2个重复,每个重复3只。上午10:00时,分别给A组-G组鳖从腹腔注射LPS(剂量分别为0,50,100,250,500,1000,5000μg/kg BW),LPS用0.5mL 0.7%NaCl配制成注射液。下午3:00时,将各组鳖断头取血,用0.5%肝素钠抗凝,2000r/min离心10min制备血浆。用放射免疫法测定血浆COR含量。结果表明:E组(500μg/kg BW)血浆COR水平极显著高于A组(对照组)、B组(50μg/kg BW)和C组(100μg/kg BW)(P＜0.01),显著高于D组(250μg/kg BW)(P＜0.05)。试验二,选择中华鳖30只(329.66±23.98)g,随机分成5组(A组-E组),每组设2个重复,每个重复3只。上午10:00时,A组(对照组)注射0.7%NaCl,B组-E组(试验组)分别注射500μg LPS/kg BW,注射剂量均为0.5mL。分别于注射当日下午3:00时A组、B组),注射后1 d(C组)、4 d(D组)和7 d(E组)的下午3:00时断头取血,制各血浆。用放射免疫法测定各血浆样品COR、TNF-α、IL-1β和IL-6的含量;用免疫浊度法测定血浆样品IgM、IgG和IgA的含量。结果表明:给中华鳖成鳖腹腔注射500μg LPS/kg BW 5h时,血浆COR、TNF-α和IL-1β的含量显著或极显著地高于对照组(P_(COR)＜0.01,P_(TNF-α)＜0.05,P_(IL-1β)＜0.01);注射1 d时,血浆IL-6的含量极显著高于对照组(P＜0.01);注射4 d时,血浆IgM的含量显著高于对照组(P＜0.05)。试验三,选择幼鳖54只(106.46±19.53)g,随机分成3组(A组-C组),每组设3个重复,每个重复6只。A组(对照组)注射0.7%NaCl,B组注射500μg LPS/kg BW,C组注射1 000μg LPS/kg BW。LPS用0.2mL 0.7%NaCl配制成注射液。试验2周后测试各组鳖的增重率和饲料系数。结果表明:对照组和500μg LPS/kg BW组幼鳖的增重率、饲料系数未出现显著差异,饲养过程中亦未观察到明显的行为变化。但1000μg LPS/kg BW组幼鳖的摄食能力减弱,增重率极显著降低(P＜0.01),且注射处理后反应强烈,相互咬噬裙边现象严重。综上,通过对不同剂量的LPS处理5 h时中华鳖成鳖血浆COR含量变化规律的观察(试验一),对注射500μg LPS/kg BW后不同时段成鳖血浆COR、TNF-a、IL-1β、IL-6、IgM、IgG和IgA含量变化的比较(试验二),以及对比注射不同剂量LPS后幼鳖摄食、生长和行为学上的变化(试验三),可见腹腔注射500μgLPS/kg BW能诱导中华鳖成鳖产生免疫应激反应;腹腔注射1000μgLPS/kg BW可显著抑制幼鳖生长;血浆COR含量的变化可作为监测中华鳖免疫应激反应的重要指标之一。
第二章,Gln缓解中华鳖免疫应激反应机制的研究。分设三个试验,即试验四至试验六。试验四,选择中华鳖60只(317.86±22.57)g,随机分为3组,A组设4个重复,每个重复3只,B、C组设8个重复,每个重复3只。上午10:00时,A组注射0.7%NaCl,B组注射500μg LPS/Kg BW,C组注射500μg LPS+100mgGln/Kg BW。LPS和Gln用0.5mL 0.7%NaCl配制成注射液。分别于注射当日(A组-C组)、注射后1 d(A组-B组)、4 d(A组-B组)和7 d(A组-C组)的下午3:00时从各组随机取2个重复6只鳖断头取血,制备血浆;解剖取相同部位肝脏组织20 g。检测不同时段(5h,1d,4d,7d)血浆COR、TNF-α、IL-1β、IL-6、GSH、GSH-PX、MDA以及肝脏PRO、GSH、GSH-PX、MDA含量的变化。结果表明:注射LPS后,血浆中COR、TNF-α、IL-1β、IL-6、MDA和肝脏组织PRO、MDA含量在相应时段显著或极显著升高(P＜0.01),而血浆GSH含量显著降低(P＜0.05);补充Gln不仅缓解了血浆COR、TNF-α、IL-1β和IL-6等的上升趋势,减缓了血浆GSH的下降,而且使肝脏MDA水平极显著降低。提示Gln可通过抑制有关细胞因子(如TNF-α、IL-1β、IL-6)和糖皮质激素(如COR)的分泌,以及增强机体的抗氧化能力而对免疫应激的中华鳖产生保护作用。试验五,选择中华鳖72只(332.35±13.75)g,随机分成4组(A组-D组),每组设6个重复,每个重复3只。A组(对照组)喂基础饲料,1周后腹腔注射0.7%NaCl;B组喂基础饲料,1周后腹腔注射500 ug LPS/kg BW;C组在基础饲料中添加2%Gln,1周后腹腔注射500 ug LPS/kgBW;D组在基础饲料中添加2%Gln,1周后腹腔注射0.7%NaCl。上午10:00时注射,注射剂量均为0.5mL。分别于注射处理后的第4d、11d和25d下午3:00从各组随机取2个重复共6只鳖称重,全身灭菌,断头取血。取表层淋巴细胞检测CD4~+/CD8~+和淋巴细胞转化率;取血浆测定IgM,IgG,lea和溶菌酶含量;解剖取小肠15cm(自幽门始),用1.5 mL0.7%NaCl反复灌洗,3000r/min离心10min,取上清液检测S-IgA。结果表明:免疫应激极显著提高中华鳖成鳖血浆IgM水平(P＜0.01),补充Gln可使LPS诱导的血浆IgM水平显著降低(P＜0.01);免疫应激抑制了中华鳖的细胞免疫功能(P＜0.05),补充Gln能缓解免疫应激对中华鳖细胞免疫功能的抑制(P＜0.05);补充Gln能显著促进肠道S-IgA的分泌(P＜0.01)。试验六,选择中华鳖36只(318.53±10.26)g,随机分成6组(A-F组),每组设2个重复,每个重复3只。A组注射0.7%NaCl;B组注射500μgLPS/kg BW;C组注射100mgGln/kgBW;D组注射100mgGln+8mgQ/kgBW;E组注射500μgLPS+100mgGln/kgBW;F组注射500μgLPS+100mgGln+8mgQ/kgBW。LPS、Gln和Q用0.5mL 0.7%NaCl配制成注射液。上午10:00时注射,下午4:00时分别取脾脏、肝脏和肠道组织,采用免疫组化法检测HSP70的表达。取血浆测定各组COR含量的变化。结果表明:注射500 ug LPS/kg BW显著促进了中华鳖肝脏和脾脏组织HSP70的合成,而注射100 mg Gln/kg BW或8 mg Q/kg BW抑制了中华鳖肠道、肝脏和脾脏组织HSP70的表达;Gln对免疫应激中华鳖的保护作用,与其诱导机体组织HSP70的表达无关。综合试验四、试验五和试验六的结果,表明Gln可通过抑制炎性细胞因子(如TNF-α、IL-1β、IL-6)和糖皮质激素(如COR)的分泌、调节免疫功能以及增强机体的抗氧化能力等途径对免疫应激中华鳖产生保护作用。
第三章,Gln/LPS对中华鳖生产性能的影响。设计一个试验,即试验七,选择中华鳖72只(101.92±17.53)g,随机分成4组(A-D组),每组设6个重复,每个重复3只。A、B组用基础饲料饲喂;C、D组饲料中添加2%Gln。第8d B、C组注射1000ugLPS/kg BW,A、D组注射0.7%NaCl。第51d进行同样处理。第95d检测各组鳖的增重率、相对生长率、饲料系数,脂体比、裙边比、肌肉的水分、粗蛋白、粗脂肪和氨基酸含量,取小肠、肝脏和脾脏切片,、观察其组织结构变化。结果表明:腹腔注射1000 ug LPS/kg BW,使幼鳖肠绒毛发育受损,肝细胞萎缩变性,显著抑制其生长,并降低肌肉品质。添加2%Gln,能促进肠道发育,显著促进生长,阻止免疫应激对中华鳖肝脏的损伤,改善免疫应激导致的中华鳖生产性能的下降。
Trionyx sinensis,an old secondary aquatic reptiles,are of research importance. Since 1980s,the increasing market demand has stimulated the rapid development of Trionyx sinensis breeding.Trionyx sinensis is always under the state of stress under the impact of various kinds of environment,human management and pathogeny,so that its ability to resist disease and muscle quality are reduced.Gln is a conditionally necessary amino acid,which has not only nutrition,but also important pharmacological action.The thesis sets up the mode of the immunological stress in Trionyx sinensis by using E.coli LPS,and makes an on-the-spot investigation into the protection and mechanism of Gln on internal secretion of the mechanism,ability to resist oxidation,immunologic function, and production performance of the immunological stressed Trionyx sinensis by injecting and adding Gln in the fodder.In order to profoundly probe into immunological stress mechanism,the application of Gln to strengthen the immunological protection lays a theoretical foundation for the healthy cultivation of Trionyx sinensis.
In the thesis,seven tests are set for the research on easing of immunological stress mechanism in Trionyx sinensis.
Forty-two healthy Trionyx sinensises(339.46±25.54)g whose weight is close to each other are chosen in test one and divided into seven groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.Each Trionyx sinensis was respectively injected into different doses of LPS from abdominal cavity,and heads were cut to get plasma after 5h. COR content in the blood plasma was measured using radioimmunoassay.It is proved that the plasma COR level content in group E is strikingly higher than that in group A(comparison group),B(50μg/kg BW) and C(100μg/kg BW)(P＜0.01),and is much higher than that in group D(250μg/kg BW)(P＜0.05).Thirty healthy Trionyx sinensises (329.66±23.98)g are chosen in test two and divided into five groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.We injected 0.7%NaCl in the comparison group and 500μgLPS/kgBW in each test group respectively,and cut their heads to get plasma at the same time after 5h、1d、4d and 7d.We measured COR、TNF-α、IL-1βand IL-6 content in each sample blood plasma using radio-immunoassay, and IgM、IgG and IgA content in the sample blood plasma using immunoturbidimetery.It is proved that when injected 500μgLPS/kgBW in the abdominal cavity of mature Trionyx sinensis for five hours,the COR、TNF-αand IL-1βcontent is much higher or strikingly higher than that in the comparison group(P_(COR)＜0.01,P_(TNF-α)＜0.05,P_(IL-1β)＜0.01),if injected for 1d,IL-6 content in the blood plasma is strikingly higher than that in the comparison group(P＜0.01),if injected for 4d,the IgM content in the blood plasma is much higher than that in the comparison group(P＜0.05).Fifty-four young Trionyx sinensises(106.46±19.53) g are chosen in test three,and divided into three groups at random,with 3 subgroups and 6 Trionyx sinensis in each subgroup.They were respectively injected into 0.7%NaCl,500μgLPS/kgBW and 1000μgLPS/kgBW.We tested weight gain ratio(WGR) and feed conversion ratio(FCR) of each group after two week's test.It is proved that there is neither obvious difference in WGR and FCR of young Trionyx sinensis in comparison group and group 500μg LPS/kg BW,nor obvious behavior difference in the process of feeding.However,young Trionyx sinensis in group1000μgLPS/kg BW has weakened food intake ability and remarkably decreased WGR(P＜0.01).Furthermore, there is strong reaction,frequent aggression of mutual biting skirt lateral after injection.In conclusion,through COR content change in the blood plasma of mature Trionyx sinensis by injecting different doses of LPS after 5h(test one),change of the COR、TNF-a、IL-1β、IL-6、IgM、IgG and IgA content in th blood plasma at different periods after injection of 500μg LPS/kg BW(test two),and changes in the food intake,growth and behavioristics of young Trionyx sinensis after injection of different doses of LPS(test three),the injection of 500μgLPS/kg BW into the abdominal cavity could cause immunological stress reaction in mature Trionyx sinensis,while injection of 1000μg LPS/kg BW into the abdominal cavity could inhibit its growth.in young Trionyx sinensises and COR content in the blood plasma could be regarded as one significant index to supervise the immunological stress reaction of Trionyx sinensis.
Sixty Trionyx sinensises(317.86±22.57) g are chosen in test four and divided into three groups at random,with 4 subgroups and 3 Trionyx sinensis in each subgroup in group A,with 8 subgroups and 3 Trionyx sinensis in each subgroup in group B and C.group A,B and C were respectively injected 0.7%NaCl、500μgLPS/Kg BW、500μg LPS+100mgGln /Kg BW into abdonamal cavity to inspect the content variation of COR、TNF-α、IL-1β、IL-6、GSH、GSH-PX、MDA in blood plasma and protein、GSH、GSH-PX、MDA in lever at different times.The result indicates that after injection of LPS,the content of COR、TNF-α、IL-1β、IL-6、MDA in blood plasma and PRO、MDA in the liver organization strikingly rise at the corresponding period(P＜0.01),while the GSH content in blood plasma remarkably lower(P＜0.05).The supplement of Gin not only eases the rising trend of COR、TNF-α、IL-1βand IL-6,the decrease of GSH in the blood plasma,but also prominently lowers the MDA level of liver.Gln could protect the immunological stressed Trionyx sinensis by restraining the secretion of concerned cell factors such as TNF-α、IL-1β、IL-6,glucocorticoid(COR) and improving the oxidatin resistance ability. Seventy-two Trionyx sinensises(332.35±13.75) g are chosen in test five and divided into four groups at random,with 6 subgroups and 3 Trionyx sinensis in each subgroup.Group A and B are fed with basal feed,group C and D are fed with basal feed plus 2%Gln.After being fed for one week,group B and C are injected into 500ugLPS/kgBW,group A and D are injected into 0.7%NaCl.4d,11d and 25d After injection the content of CD_4~+/CD_8~+ in peripheral blood,transformation ratio of T lymph,IgM、IgG、IgA in the blood plasma, muramidase and S-IgA content in intestinal canal were examined.The result proves that the immunological stress caused by LPS strikingly improves the IgM level(P＜0.01) in the blood of Trionyx sinensis.Supplement of Gln not only improves the immunologic function of cells and intestinal canal,but also prohibits the rising of IgM level in the blood plasma conducted by LPS.Thirty-six Trionyx sinensises(318.53±10.26)g are chosen in test six and divided into six groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.We used LPS、Gln and Q to treatt them.After 6h,we got spleen,liver and intestinal canal organization and inspected the expression of HSP70 by immunochemistry.At the same time,we inspected the COR content variety of each group. It is proved that injection of 500ugLPS/kgBW remarkably improves the expression of HSP70 in spleen and liver organization of Trionyx sinensis,while injection of 100mgGln/ kgBW or 8mgQ/kgBW inhibits the expression of HSP70 in intestinal canal,spleen and liver organization.To sum up,Gln could protect the immunological stressed Trionyx sinensis by regulating the immunological function,inhibiting the secretion of inflammatory cell factor and improving the oxidation resistance ability,while the protection might have nothing to do with the expression of HSP70.
Seventy-two Trionyx sinensises(101.92±17.53) g are chosen in test seven and divided into four groups at random,with 6 subgroups and 3 Trionyx sinensis in each subgroup.Group A and B are fed with basal feed,and group C and D are fed with basal feed plus 2%Gln.On the eighth day,group B and C are injected into 1000ugLPS/kg BW and 0.7%NaCl is injected into group A and D.On 51th day,they are under the same treatment,on 95th day,we inspected the weight gain ratio,comparison growth ratio, feed conversion ratio,the content of body fat ratio,body skirt ratio,muscle moisture, coarse protein,coarse fat and amino acid,and got small intestine,liver and spleen to observe their structure variation.It is proved that injection of 1000ugLPS/kgBW caused the damage of the villus growth of the intestine,the shrinkage and degeneration of liver cells,which strikingly inhibits the growth of the young Trionyx sinensis and lowers the quality.Supplement of 2%Gln could improve the growth of intestine,remarkably enhance the growth,restrain the damage of immunological stress to the liver of Trionyx sinensis,and improve the production performance of Trionyx sinensis reduceded by immunological stress.
引文
[1]Beisel,W.R.1977.Magnitude of host nutritional response to infection.Am.J.Clin.Nutr.30:1236-1247.
[2]Van Heugten,E.,J.W.Spears,and M.TCofey.1994.The efect of dietary protein on performance and immune response in weaning pigs subjected to an inflammatory challenge.J.Anim.Sci.72:2661-2669.
[3]Webel,D.M.,R.W Johnson and D.H.B aker.1998.L ipopolysaccharide-induced reductions in food intake do not Decrease the eficiency of lysine and threonine utilization for protein accretion.J.Nutr.128:1760-1766
[4]Johnson,R.W.1997.Inhibition of growth by pro-inflammatory cytokines:an integrated view.J.A nim.Sci.75:1244.
[5]Andus,T,J.Bauer,and W Gerok.1991.Efects of cytokines on the liver.Hepatology.13:364-375.
[6]Williams,N.H,T.S.Stahly,and D.R.Zimmerman.1997.Efect of chronic system activation on the rate,efiency,and Composition of growth and lysine needs of pigs fed from 6 to 27 kg.J.Anim.Sci.75:2463-2471.
[7]王道尊,汤峥嵘.中华鳖生化组成的分析.水生生物学报,1997,21(4):299-305
[8]汤峥嵘,王道尊.中华鳖生化组成的分析:Ⅲ.肌肉氨基酸的组成.水生生物学报,1998,22(4):307-313
[9]张轩杰,陈平.中华鳖氨基酸组成的反相高效液相色谱分析.湖南师范大学自然科学学报,1997,20(1):72-76
[10]占秀安,许梓荣.中华鳖肉脂品质的研究.浙江大学学报:农业与生命科学版,2000,26(4):457-460
[11]王道尊,汤峥嵘.中华鳖生化组成的分析:Ⅱ.背甲,肌肉中矿物元素的组成.水生生物学报,1998,2(22):106-111
[12]刘桂林,聂向庭.甲鱼脂肪酸和微量元素含量的分析及其抗氧化作用研究.营养学,2000,22(4):325-327
[13]吴惠仙,王淑霞.中华鳖疾病研究进展.水产科学,1999,18(5):38-41
[14]占秀安.野生鳖与养殖鳖肌肉营养,保健价值的比较研究.大连水产学院学报,2000,15(2):108-112
[15]占秀安.池塘鳖与温室鳖体组成和生化组成的比较研究.大连水产学院学报,2001,(4):269-273
[16]朱秋华,钱国英.仿生鳖与温室鳖的营养成分比较.特产研究,2001,23(3):39-42
[17]Winduller H G.Glutamine utilization by the small intestine[J].Adv Enzymol,1982;53:210
[18]ChenK,Okuma T,Okamura K et al.Glutamine-supplemented parenteral nutrition improves gut mucosa integrity and function in endotoxemic rats[J].JPEN,1994;18(2):167
[19]杨俊涛.谷氨酰胺与免疫细胞[J].国外医学创伤与基本问题分册,1997;18:6
[20]Crawford J,Cohen HJ.The essential role of L-glutamine in lymphocyte diferentiation in vitro[J].J Cell Physiol,1985;124(2):310-315
[21]黄耀凌,邹思湘.谷氨酰胺的抗疲劳生化机制研究[J].南京农业大学学报,2001,24(2):87-89
[22]Yoshida S,LeskiwM J,SchuterM D et al.Efectof total parenteral nutrition,systemic sepsis,and glutamine on gut Mucosa in rats[J].Ann J Physiol,1992,263:E368
[23]Jacobs DO,Evans DA,Mealy K et al.Combined efects of glutamine and epidermal growth factor on the rat in testine[J].Surgery,1988;104:358
[24]于健春,蒋朱明,李德敏.谷氨酰胺调节谷胱甘肽的代谢,保护肝脏机理的基础和临床研究[J].肠外与肠内营养,1996;3(2):S32
[25]张军民,王连递,高振川等.日粮中添加谷氨酰胺对早期断奶仔猪抗氧化能力的影响[J].畜牧兽医学报,2002,33(2):105-109
[26]Dritz,S.S.,K.Q.O wen and R.D.Goodband.1996.Influence of ipopolysaccharideinduced immune challenge and diet c omplexity on growth performance and acutephase protein production in segregated early-weaned pigs.J.Anim.Sci.74:1620-1628.
[27]Klasing K C,Barnes D M.Decreased amino acid requirements of growing Chicks due to immunologic stress.1988,Nutr,118:1158-1164
[28]Thiel-Cooper R L,Parrish F C Jr,Sparks J C,et al.Conjugated linoleic acid changes swine performance andcarcass composition[J].J Anim Sci,2001,79:18211828
[29]Dugan M E R,Aalhus J L,Jeremiah L E.et al.The effects of feeding conjugated linoleic acid on subsequent pork quality[J].Can J Anim Sci,1999,79:45-51
[30]Klasing K C,Laurin D E,Peng R K,et al.Immunologically mediated growth depression in chicks:Influence of feed intake,corticosterone and interleukin-1[J].J Nutr,1987,117:1629-1637
[31]Webel D M,Finck B N,et al.Time course of increased plasma cytokines cortisol and urean itrogen in pigs following intraperitoneal injection of lipopoly saccharide.1997,nim.Sci.75:1514-1520
[32]赖长华,尹靖东,李德发,等.共轭亚油酸对免疫应激仔猪生长抑制的缓解作用.中国畜牧杂志,2005,41(2):6-9
[33]宋林生,邱丽华,蔡中华,等.花鲈肿瘤坏死因子基因及其在LPS和病毒刺激下表达的研究.High TechnOlogy Letters.2004,14(11):81-86
[34]Besedovsky,H,D.R.Adriana,S.Ernst and C.A.Dinarello.1986.nununoregulatory feedback between in terleukin-1 and glucocorticoid hormones.J.Science.233:652-654.
[35]Carroll,J.A.,K.J.Touchete,R.L.Matteri,C.J.Dyer,and G.L.Allee.2002.Efect of spraydried plasma and lipopolySaccharide exposure on weaned pigs:Ⅱ.Efectson the hypothalamicpituitaryadrenal axis of weaned pigs.J.Anim.Sci.80:502-509.
[36]Parrot,R.F.,S.V.Vellucci,and J.A.Goode.1997.Interrelated adrenocortical and neurohypo-Phy sical responses Associated with fever in endotoxin-treated pigs.Am.J.Physiol.273:R1046-R1052.
[37]Ruane N M,Nolan D T,Rotllant J,et al.Experimental exposure of rainbow trout Oneorhynch sykiss(Walbaum) to the infective stages of the sea louse Lepeophtheirus saimonis(Kroyer)influences the physiological response to an acute stressor.Fish & Shell fish Immunology,2000,10:451-463.
[38]Fong,Y,L.L.Moldawer and F.Lowry.1989.Cachectin/TNF or IL-1 induces cachexia with redistribution of body Proteins.Am.J.Physiol.R659-R665.
[39]李建涛,蔡辉益,刘国华等.免疫应激对肉仔鸡采食量、能量及氮消化率和内源氮损失的影响.河北农业大学学报,2005,28(6):90-94
[40]Klasing,K C,and B.J.Johnstone.1991.Monokines in growth and development.Poutr.Sci.70:1781-1789.
[41]Jepson M.M,J.M.Pell,P.C.Bates,and D.J.Millward.1986.The efects of endotoxaemia on protein metabolism in skeletal muscle and liver of fed and fasted rats.J.Bi-ochem.235:329-336.
[42]Memon R A,Feingold K R,Grunfeld C.The efect of cytokines on intermediary Metabolism [J].Endocrinologist,1994,4:59-63
[43]Wright,K.J.,R.Balaji,C.M.HiII,S.s.Dritz,E.L.Knoppel,audJ.E.Minton.Integrated adrenal,soma-Tot-ropic,and immune responses of g owing pigs to treatment with lipopolysaccharide.J.Amim.Sci.,2000,78:1892-1899
[44]杨汉春主编.动物免疫学口[M].北京:中国农业大学出版社,2003
[45]Balaji,R.,K.J.Wright,C.M.Hil],S.S.Dritz,E.L.Knoppel,and J.E.Minton.Acute phase responses pigs challenged orally with Salmonella typhimurium.J.Anim.Sci.,2000,78:1885-1891
[46]白满喜,蒋朱明.谷氨酰胺双肽对大鼠肠切除后细菌移位的影响.中华医学杂志,1996,76(2):116-119
[47]龚时文,张明.谷氨酰胺对大鼠急性坏死性胰腺炎肠道细菌移位的影响.中国急救医学,2000,20(11):640-641
[48]刘涛,周诗其,程学慧等.外源性谷氨酰胺和谷氨酸对早期断奶仔猪肠粘膜形态、结构和小肠吸收功能及骨骼肌中DNA、RNA浓度的影响.中国兽医学报,2003,2391):62-65
[49]石慧琳,程义勇,李树田等.补充谷氨酰胺对束缚大鼠肠屏障功能的影响.营养学报,2004,26(1):31-35
[50]范志勇,王康宁,周定刚.EGF、Gln和pGRF基因质粒对早期断奶仔猪肠道发育的影响.畜牧兽医学报,2006,37(5):457-463
[51]席鹏彬,林映才,蒋宗勇等.谷氨酰胺二肽对断奶仔猪生长、免疫、抗氧化力和小肠粘膜形态的影响.动物营养学报,2007,19(2):135-141
[52]ButIer,EJ.,M.J.,Curtis,and E.G:,Harry.Escherichia coli endotoxin as a stressor in the domestic fowl.Research in veterninary Sci.,1977,23:20-23
[53]Butts,C.A.,Moughan,P.J.,Smith,W.C.Endogenous amino acid flow at the terminal ileum of the rat under conditions of peptide atimentation.J.Sci.Food Agri.,1991,55:175-187
[54]Butts,C.A.,Moughan,P.J,Smith,W.C.,Reynolds,GW.,and Garrick,D.J.The efects of food,dry matter intake on endogenous ileal amino acid excretion determined under peptide alimentation in the 50kg liveweight pig.J.Sci.Food Agri.,1993,62:235-243
[55]Chung,T.K.,and D.H.Baker.Apparent and true amino acid digestibility of a crystalline amino acid mixture and of casein:comparison of values obtained with ileal-cannulated pigs and cecectomized cockerels.J.Anim.Sci.,1992,70:3781-3790.
[56]Chung,T.K,J.Khajareen,and D.R.Zimmerman.Efect of dietary threonine on growth performance and carcass Characteristic of broiler chickens raised at hight Emperatures.In:proceedings of the World's Poultry Congress,New Delhi,India,1996,p.145
[57]Coats,M.M.,FuIIer,r.,et al.A comparision of the growth of chickens in the gustafsson germfree paratus and in conventional environment,with and w ithout dietary supplements of penicillin.Br.J,Nutr.,1963,17:141-150
[58]Colley,C.M.,Fleck,A,Googe,A.W,Muller,B.R.,and Myers,M.A.Early time course of acute phase protein response in man,J.Clin.Path.,1983,36:203-207
[59]周吕.胃肠生理学-基础与临床(M].北京:科学出版社,1991,98-105
[60]Yagoob P,Calder PC.Glutamine requirement of proliferatin T-lymphocytes[J].Nutrition,1997,13:646-651
[61]黄耀凌,邹思湘,陈伟华.谷氨酰胺对小鼠免疫功能的影响[J].营养学报,2001,23(4):363-364
[62]杨俊涛.谷氨酰胺与免疫细胞[J].国外医学创伤与基本问题分册,1997;18:6
[63]林燕.谷氨酰胺对幼建鲤肠道功能和免疫力的影响.四川农业大学:硕士学位论文2005
[64]舒晓亮,曹翔,耿珊珊等.谷氨酰胺对严重创伤手术后患者应激反应和免疫功能的影响.同济大学学报:医学版,2005,26(6):32-34
[65]徐文萍,嵇月娥,施文艳,曹永安.补充谷氨酰胺对急性应激大鼠脑组织GSH、SOD和MDA 含量的影响.中国应用生理学杂志,2003,19(4):415-416
[66]Goldstein L,Schrock H,Cha CJ.Relationship of muscole glutamine to renal ammonia metabolism[J].Biochem Soc Trans,1980,8:509-510
[67]戴四发,金光明,王立克,柳卫国.Gln对肉用仔鸡免疫器官生长发育和成活率的影响.中国畜牧杂志.2003,039(004):25-27
[68]戴四发,王立克等.外源Gln对AA肉鸡肠道早期生长发育的影响.中国饲料.2002.000(016):12-13
[69]Yu B,You Z,Wang S.Enhancement of gut immune function by early enteral feeding enriched with L-glutamine in severe burned miniswines.Zhonghua Zheng Xing Shao shang wai Ke Za Zhi.1996,Mar;12(2):98-100.
[70]刘伟敏,刘忠臣,李新艳,唐光武.Gln肽在断奶仔猪饲料中的应用.饲料博览,2003,000(006):29-30
[71]谈定玉.大黄及谷氨酰胺对肠缺血再灌注大鼠肠道保护作用及其机理研究.[学位论文].南京中医药大学,2006
[72]王维.丙氨谷氨酰胺对大鼠急性肺损伤的影响及其机制.[学位论文].广西医科药大学,2006
[73]Balm,P.H.M,.Stress Physiologyin Animals.Sheffietd Academic Press,Sheffield,England.999.
[74]李静.猪热应激反应适宜监测指标初探[D].沈阳农业大学,2003.
[75]杨春花,黄烽.高敏C反应蛋白研究进展[J],中华风湿病学杂志,2004,8(12):755-758.
[76]计红,杨焕民,吴永魁.热应激蛋白研究进展及其应用前景[J],生物学杂志,2005,22(4):1-3.
[77]张根军.生长猪热应激反应模型及适宜监测指标筛选[D].中国农业科学研究院,2004.
[78]杨钢,席正雄,万瑜等.应激时大鼠血浆、脑、心血管及肾上腺血管紧张素Ⅱ含量的变化[J],生理学报,1993,45(5):505-509.
[79]Mahapatra MS,Mahata SK,Maiti B.Effect of stress on serotonin,norepinephrine,epinephrine and corticosterone contents in the soft-shelled turtle.Clin Exp Pharmacol Physiol.1991,18(10):719-724.
[80]Ray PP,Chaudhuri-Sengupta S,Maiti BR.Adrenocortical involvement during diverse stress in soft-Shelled turtle(Lissemys p.punctata) Bonnoterre.Indian J Exp Biol.2004,42(6):620-625.
[81]Rudy M.Ortiz:Rhonda M.Pattersonb;Charles E.Wadea;F.Michael Byersc.Effects of acute fresh water Exposure on water flux rates and osmotic responses in Kemp's ridley sea turtles (Lepidochelys.kempi) Comparative Biochemistry and Physiology.Part A:Molecular &Integrative Physiology.2000,127,(1):81-87
[82]Ray PP,Maiti BR.Adrenomedullary Hormonal and Glycemic Responses to High Ambient Temperature in the Soft-Shelled Turtle(Lissemys punctata punctata).General and Comparative Endocrinology.2001,122(1):17-22.
[83]Ray PP,Maiti BR.Interrenal responses to high ambient temperature in softshelled turtle,(Lis-semys punctata Punctata).Indian J Exp Biol.2003,41(8):880-884.
[84]Jessop,T.S.,Hamann,M.,Read,M.A.,Limpus,C.J.Evidence for a hormonal tactic maximining green Turtle ReproDuction in response to a pervasive ecological stressor.General and Comparative Endocrinology.2000,118:407-417.
[85]Gregory,L.F.,Gross,T.S.,Bolten,A.B.,Bjorndal,K.A.,Guillette,L.J.Plasma corticosterone concentrations associated with acute captivity stress in wild logger head sea turtles(Caretta caretta).General and Comparative endocrinology.1996,104:312-320.
[86]Gregory L.F.,Schmid,J.R.Stress Responses and Sexing of Wild Kemp's Ridley Sea Turtles (Lepidochelys Kempii) in the Northeastern Gulf of Mexico.General and Comparative Endocrinology.2001,124(1):66-74.
[87]William B.Cash.,Rebecca L.Holberton.,Scott S.Knight.Corticosterone Secretion in Response to Capture and Handling in Free-Living Red-Eared Slider Turtles.General and Comparative Endocrinology.1997,108(3)427-433.
[88]Ott,J.A,Mendonc,M.T.,Guyer,C,,Michener,W,K.Seasonal changes in sex and adrenal teroid hormones of gopher tortoises(Gopherus polyphemus).General and Comparative Endocrinology.2000,117(2):299-31.
[89]Zhou Xianqing,Xie Mengxia,Niu Cuijuan,Sun Ruyong.The effects of dietary vitamin C on growth,liver Vitamin C and serum cortisol in stressed and unstressed juvenile soft-shelled turtles (Pelodiscussinensis).Comparative Bioch-emistry and Physiology Part A:Molecular &Integrative Physiology.2003,135(2):263-270.
[90]Zhou,Xianqing.,Niu,Cuijuan.,Sun,Ruyong.The effects of vitamin E on antiacid stress ability in juvenile soft-shelled turtles(Pelodiscus sinensis).Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology.2004,137(4):299-305.
[91]Jessop,T.S.Modulation of the adrenocortical stress response in marine turtles(Cheloniidae):evidence for a hormone tactic maximizing reproductive investment.J.Zool.Lond.2001,254:57-65.
[92]Valverde,R.A.,Owens,D.W.,Mackenzie,D.s.,Amoss,M.s.Basal and stress-induced corticosterone in olive Ridley sea turtles(Lepidochelys olivacea) in relation to their mass nesting behavior.J.Exp.Zool.1999a,284:652-662.
[93]Valverde,R.A.,Owens,D.W.,Mackenzie,D.s.,Amoss,M.s.Adrenal responsiveness during of the clive ridley sea turtle(Lepidochelys olivacea).Am.Zool.1999b,32A,21.
[94]Jessop,T.s.,Knapp,R,Limpus,C.J.,Whittier,J.M.Dynamic endocrine responses to stress:evidence for energetic constraints and status dependence of breeding in male green turtles.Gen.Comp.Endocrinol.2002,126:59-67.
[95]Aguirre,A.A.,Balazs,G.H.,Spraker,T.R..Adrenal and hematological responses to stress in juve-nile Greenturtles(Chelonia mydas) with and without fibropappillomas.Physiol Zool.1995,68:831-854.
[96]Beatrix G Schramm.,M Casares.,Valentine A.Lance.Steroid Levels and Reproductive Cycle of the Galapagos Tortoise(Geochelone nigra) Living under Seminatural Conditions on Santa Cruz Island(Galapagos).General and Comparative Endocrinology.1999,114(1):108-120.
[97]周显青,牛翠娟,孙儒泳.黄芪和酸应激对中华鳖幼鳖血清补体C3和C4含量的影响[J],动物学研究,2002,23(2):177-180.
[98]周显青,牛翠娟,孙儒泳.饵料维生素E含量对酸应激中华鳖幼鳖血清补体C_3和C_4的影响[J],动物学研究,2003a24(6):452-456
[99]周显青,牛翠娟,孙儒泳.维生素C和酸应激对中华鳖幼鳖血清补体C_3和C_4含量的影响[J],动物学报,2003b,49(6):769-774.
[100]周显青,谢孟峡,牛翠娟,孙儒泳.维生素C和E合用对应激和非应激中华鳖幼鳖生长、肝脏维生素C和E以及血清皮质醇含量的影响[J],动物学报,2004a,50(2):158-164.
[101]陈欣然,牛翠娟,蒲丽君.慢性氨氮暴露对中华鳖生长及非特异性免疫功能韵影响[J],北京师范 大学学报:自然科学版,2006,42(3):300-304.
[102]Willmore WG,Storey KB.Multicatalytic proteinase activity in turtle liver:responses to anoxia stress and recOvery Biochem Mol Biol Int.1996,38(3):445-51.
[103]Willmore WG,Storey KB.Purification and properties of the glutathione S-transferases from the anoxiatolerant turtle(Trachemys scripta elegans).FEBS J.2005,272(14):3602-3614.
[104]Hossein Mehrani;Kenneth B.Storey.Enzymatic control of glycogenolysis during anoxic submergence in the Fresh water turtle(Trachemys scripta).The International Journal of Biochemistry & Cell Biology.1995,27(8):821-830.
[105]Bagatto,B,Henry,RP.Exercise and forced submergence in the pond slider(Trachemys scripta)and softshell turtle(Apalone ferox):influence on bimodal gas exchange,diving behaviour and blood acid-base status.Journal of Experimental Biology.1999,202(3):267-278.
[106]Jackson DC,Warburton SJ,Meinertz EA,Lawler RG,Wasser JS.The effect of prolonged anoxia at 3 degrees C on tissue high energy phosphates and phosphodiesters in turtles:a 31P-NMR study.J Comp Physiol(B);1995,165(1):77-84.
[107]Wasser JS,Heisler N.Effects of anoxia on intracellular free Ca~(2+) in isolated cardiomyocytcs from turtles.Comp Biochem Physiol A Physiol.1997,116(4):305-12.
[108]Bobb VT,Jackson DC.Effect of graded hypoxic and acidotic stress on contractile force of heart muscle from ypoxiatolerant and hypoxia-intolerant turtles.J Exp Zoolog A Comp Exp Biol.2005,303(5):345-53.
[109]Attilio Arillo;Emilio Balletto;Maria A.Cherchi.Quantitative variations of some compounds of the T.C.A.-cycle in specimens of Testudo hermanni grnelin after cxposure to hyperthermic conditions.Journal of Thermal Biology 1997,2(3):131-133.
[110]Michel Cahanac,Cecile Bernieri..Behavioral rise in body temperature and achycardia by handling of a Turtle(Clemmys insculpta).Behavioural Processes.2000,49(2):61-68.
[111]Rotermund A.J;Privitera C A..The effects of induced cold torpor on ATPase activity of the turtle,Pseudemys(chrysemys) picta.Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology.1972,41(3):511-520.
[112]Emilio Balletto,Maria A.Cherchi,Federico Melodia,Paola Mensi.Temperaturcdependent thyroid hormone Bios-ynthesis in Testudo horsfieldi Gray.General and Comparative Endocrinology.1979,39(4):548-551.
[113]Baccari GC,Minucci S,Di Matteo L.The orbital glands of the terrapin Pseudemys scripta in response to osmotic stress:a light and electron microscope study.J Anat.1993,183(1):21-33.
[114]周显青,牛翠娟,李庆芬等.光照强度对中华鳖稚鳖摄食和生长的影响[J],动物学报,1998a,44(2):157-161.
[115]周显青,牛翠娟,李庆芬等.光照度对中华鳖稚鳖能量转换的影响[J],北京师范大学学报(自然科学版),1998b,34(2):248-251.
[116]周显青,牛翠娟,李庆芬.光周期对中华鳖稚鳖摄食,生长和能量转换的影响[J],生态学报,1999a(3):383-387.
[117]Guillette Jr.,L.J.,Cree,A.,Rooney,A.A.biology of stress:interactions with reproduction,immunology and Inter mediary etabolism,in:Warwick,C.,Frye,F.L,Murphy,J.B.(eds),Health and welfare of Captive Reptiles,Chapman and hall,London,pp.1995,32-81.
[118]Mahmoud,I.Y;Licht,Paul.Seasonal Changes in Gonadal Activity and the effects of Stress on Reproductive Hormones in the Common Snapping Turtle(Chelydra serpentina).General and Comparative Endocrinology.1997,107(3):359-372
[119]Licht P,Breitenbaeh GL,Congdon JD.5Seasonal cycles in testicular activity,gonadotropin,and thyroxine in the painted turtle(Chrysemys picta) under natural conditions.Gen Comp Endocrinol.1985,59(1):130-9.
[120]Huot-Daubremont C.;Bradshaw S.D.;Bradshaw F.J.;Kuchling G;Grenot C.J.variation of plasma sex steroid concentrations in wild and captive populations of hermann's tortoise(Testudo hermanni hermanni) in Southern France.General and Comparative Endocrinology.2003,130(3):299-307.
[121]Jessop,T.S.,Limpus,C.J.,Whittier,J.M.Plasma steroid interactions during highdensity green turtle nesting and associated disturbance.Gen.Comp.Endocrinol.1999,115,90-100.
[122]Prassack SL,Bagatto B,Henry RP.Effects of temperature and aquatic P(O_2) on the physiology and behaviour of Apalone ferox and Chrysemys picta.The Journal of Experimental Biology.2001,204,2185-2195.
[123]Jon P Costanzo;Jaequeline D Litzgus;Richard E LeeJr.Behavioral responses of hatchling painted turtles(ChryseMys picta) and snapping turtles(Chelydra serpentina) at subzero temperatures.Journal of Thermal Biology.1999,24(3):161-166.
[124]周显青,牛翠娟,李庆芬.光照对水生动物行为的影响[J],动物学杂,1999b,34(2):45-48.
[125]Richard W.Brill;George H.Balazs;Kim N.Holland;Randolph K.C.Chang;Scott Sullivan;John C.George.Daily movements,habitat use,and submergence intervals of normal and.tumor-bearing juvenile green turtles(Chelonia mydas L).within a foraging area in the Hawaiian islands.Journal of Experimental Marine Biology and Ecology.1995,185(2):203-218.
[126]王军,赵景宏,粟永萍.糖皮质激素受体调控与创伤应激紊乱关系的研究进展[J],中华创伤杂志,2001,17(8):508-510.
[127]Rice M.E;Forman R.Chen E;B.T;Avshalumov M.V;Cragg S.J;Drew K.L.Brain antio-xidant regulation in mammals and anoxia-tolerant reptiles:balanced for neuroprotection and neuromodulation.Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology.2002,133(4):515-525.
[128]周显青,牛翠娟,孙儒泳.维生素E对中华鳖幼鳖生长、肝脏维生素E以及血清皮质醇含量的影响[J],动物学报,2003c,49(1):40-44.
[129]周显青,牛翠娟,孙儒泳.维生素C和E合用对中华鳖幼鳖非特异性免疫功能的影响(J),水生生物学报,2004b,28(4):356-360.
[130]周显青,牛翠娟,孙儒泳.黄芪对中华鳖免疫和抗酸应激能力的影响[J],水生生物学报,2003d,27(1):110-112.
[131]肖明松,王志耕,崔峰,等.果寡糖和糖萜素对中华鳖生长性能及免疫功能的影响.水利渔业,2004,24(3):20-22
[132]刘宗英,姚鹃,陈昌福,等.高活性干酵母对中华鳖非特异性免疫功能和抗病力的影响.华中农业大学学报,2005,24(2):192-196
[133]汪成竹,姚鹃,吴凡,等.免疫多糖(酵母细胞壁)对中华鳖非特异性免疫功能的影响.华中农业大学学报,2006,25(4):421-425
[134]覃川杰,汪成竹,陈晓辉等.茯苓多糖对中华鳖非特异性免疫功能的免疫调节作用.淡水渔业,2006,36(6):40-43
[66]王亭亭,蔡完其.饲料中添加花粉和酵母硒对中华鳖幼鳖生长和非特异性免疫功能的影响.上海水产大学学报,2005,14(2):97-102
[135]宋超,牛翠娟,陈欣然.壳聚糖对急性氨氮胁迫下中华鳖稚鳖非特异性免疫反应的影响.动物学报,2007,53(2)270-277
[136]王亭亭,蔡完其.饲料中添加花粉和酵母硒对中华鳖幼鳖生长和非特异性免疫功能的影响.上海水产大学学报,2005,14(2):97-102
[137]刘至治,蔡完其,季高华,等.几种免疫增强剂对中华鳖红细胞数量及免疫功能的影响.上海水产大学学报,2006,15(1):1-15
[138]赵燕,代兵,李传普等.半胱胺对中华鳖生长性能和非特异性免疫功能的影响研究.动物营养学报,2007,19(3)305-310
[139]陈昌福.柱状嗜纤维菌脂多糖对鲤不同组织中离体培养淋巴细胞转化率的影响.华中农业大学学报,1998,17(1):66-70
[140]陈昌福,吴志新.三种爱得华氏菌脂多糖对日本鳗鲡免疫原性的比较.水生生物学报,1998,22,增刊:126-131
[141]陈昌福,楠田理一.三种鳜对柱状嗜纤维菌脂多糖免疫应答的比较研究.华中.农业大学学报,1999,18(3):252-255
[142]陈昌福,曾妍雄,楠田理一.接种不同种类病原菌脂多糖对翘嘴鳜细菌性败血症的免疫保护力比较.华中农业大学学报,2000,19(4):377-380
[143]陈昌福,罗宇良,曾妍雄.不同血清型柱状嗜纤维菌脂多糖蛋白质成分及其对鳜免疫原性影响的研究.华中农业大学学报,2000,19(5):483-486
[144]曾慷,何建国,王晓红,翁少虹,左涛.传染性脾肾坏死病毒,Con A和LPS对鳜的头肾淋巴细胞转化的影响.水产学报,2001,25(5):464-468
[145]黄辨非,陈超然,毛之娟,陈昌福.同腹异育银卿对鱼类3种致病弧菌脂多糖的免疫应答.华中农业大学学报,2002,21(1):59-63
[146]陈昌福,陈超然.鱼类三种致病菌的粗脂多糖对异育银卿的免疫原性.水生生物学报,2002, 26(5):483-488
[147]Trapnell,JE.Pathophysiology of acute pancreatitis.World J Surg,1981,5(3):319-327
[148]田文霞,徐彤,郑明学等.实验性免疫应激雏鸡免疫器官抗体生成细胞的动态变化研究.山西农业大学学报,2000,(3):232-235
[149]徐彤,高文伟,任家琰等.实验性免疫应激对雏鸡血液激素与免疫功能的影响.华北农学报,2002,17(1):114-118
[150]Johnson R W.Inhibition of growth by proinflammatory cytokines:An intergrated view.Journal of Animal Science,1997,75:1244-1255
[151]林清华.免疫学实验[M].武汉:武汉大学出版社,1995:188-195
[152]Knodel M,Kuss AW,Berberich,et al.Blimp-1 over-expression abrogates IL-4 and CD40 mediated of terminal B cell differentiation but arrests isotype switching[J].Eur J Immunol,2001,31:1972-1980
[153]简纪常,吴婷婷,杨弘,董在杰,曹莹.中华鳖体液免疫应答的初步研究.中国水产,1998,12(4):6-10
[154]简纪常.中华鳖血清中抗体含量的研究.动物学研究,2001,22(2):163-165
[155]Sanders M M,Ken C.Glutamine is a powerful efector of heat shock protein expression in Drosophila Kccens[J].J Cell Physiol,1992,146:180.
[156]W.ischaneyer P E,Kahana M D,Wolfson R,et al.Glutamine induces heat shock protein and protects against endotoxin shock in the rat[J].J Appl Physiol,2001,90,2403.
[157]张丽英.饲料分析及饲料质量检测技术(第二版)[M].北京:中国农业大学出版社,2003:56:59
[158]陈静,刘显军,边连全,石娇.谷氨酰胺对早期断奶仔猪生长性能和免疫性能的影响.西北农业学报,2006,15(4):58-62
[159]高春生,王艳玲,杨国宇,等.谷氨酰胺(Gln)对早期断奶仔猪生长性能和腹泻的影响.中国农学通报.2006年22(4):12-14
[160]范志勇,王康宁,周定刚.EGF、Gln和pGRF基因质粒对早期断奶仔猪生产性能、免疫功能及相关激素水平的影响.畜牧兽医学报,2005,36(9):893-899
[161]果磊,贺光照,黄崇本.谷氨酰胺对烧伤大鼠肠道粘膜损伤的保护作用.重庆医科大学学报,2002,27(4):403-406.
[162]陈秋生,聂其灼.中华鳖脾脏白髓的组织结构与组织化学特性.甘肃农业大学学报,1993,28(3):220-223
[163]陈秋生,聂其灼.中华鳖脾脏的显微与亚显微结构研究.南京农业大学学报,1995,18(4):91-97
[164]赵万鹏,别立洁.鳖脾脏的正常组织学和病理组织学.信阳师范学院学报(自然科学版),2003,16(1):45-48
[2]Van Heugten,E.,J.W.Spears,and M.TCofey.1994.The efect of dietary protein on performance and immune response in weaning pigs subjected to an inflammatory challenge.J.Anim.Sci.72:2661-2669.
[3]Webel,D.M.,R.W Johnson and D.H.B aker.1998.L ipopolysaccharide-induced reductions in food intake do not Decrease the eficiency of lysine and threonine utilization for protein accretion.J.Nutr.128:1760-1766
[4]Johnson,R.W.1997.Inhibition of growth by pro-inflammatory cytokines:an integrated view.J.A nim.Sci.75:1244.
[5]Andus,T,J.Bauer,and W Gerok.1991.Efects of cytokines on the liver.Hepatology.13:364-375.
[6]Williams,N.H,T.S.Stahly,and D.R.Zimmerman.1997.Efect of chronic system activation on the rate,efiency,and Composition of growth and lysine needs of pigs fed from 6 to 27 kg.J.Anim.Sci.75:2463-2471.
[7]王道尊,汤峥嵘.中华鳖生化组成的分析.水生生物学报,1997,21(4):299-305
[8]汤峥嵘,王道尊.中华鳖生化组成的分析:Ⅲ.肌肉氨基酸的组成.水生生物学报,1998,22(4):307-313
[9]张轩杰,陈平.中华鳖氨基酸组成的反相高效液相色谱分析.湖南师范大学自然科学学报,1997,20(1):72-76
[10]占秀安,许梓荣.中华鳖肉脂品质的研究.浙江大学学报:农业与生命科学版,2000,26(4):457-460
[11]王道尊,汤峥嵘.中华鳖生化组成的分析:Ⅱ.背甲,肌肉中矿物元素的组成.水生生物学报,1998,2(22):106-111
[12]刘桂林,聂向庭.甲鱼脂肪酸和微量元素含量的分析及其抗氧化作用研究.营养学,2000,22(4):325-327
[13]吴惠仙,王淑霞.中华鳖疾病研究进展.水产科学,1999,18(5):38-41
[14]占秀安.野生鳖与养殖鳖肌肉营养,保健价值的比较研究.大连水产学院学报,2000,15(2):108-112
[15]占秀安.池塘鳖与温室鳖体组成和生化组成的比较研究.大连水产学院学报,2001,(4):269-273
[16]朱秋华,钱国英.仿生鳖与温室鳖的营养成分比较.特产研究,2001,23(3):39-42
[17]Winduller H G.Glutamine utilization by the small intestine[J].Adv Enzymol,1982;53:210
[18]ChenK,Okuma T,Okamura K et al.Glutamine-supplemented parenteral nutrition improves gut mucosa integrity and function in endotoxemic rats[J].JPEN,1994;18(2):167
[19]杨俊涛.谷氨酰胺与免疫细胞[J].国外医学创伤与基本问题分册,1997;18:6
[20]Crawford J,Cohen HJ.The essential role of L-glutamine in lymphocyte diferentiation in vitro[J].J Cell Physiol,1985;124(2):310-315
[21]黄耀凌,邹思湘.谷氨酰胺的抗疲劳生化机制研究[J].南京农业大学学报,2001,24(2):87-89
[22]Yoshida S,LeskiwM J,SchuterM D et al.Efectof total parenteral nutrition,systemic sepsis,and glutamine on gut Mucosa in rats[J].Ann J Physiol,1992,263:E368
[23]Jacobs DO,Evans DA,Mealy K et al.Combined efects of glutamine and epidermal growth factor on the rat in testine[J].Surgery,1988;104:358
[24]于健春,蒋朱明,李德敏.谷氨酰胺调节谷胱甘肽的代谢,保护肝脏机理的基础和临床研究[J].肠外与肠内营养,1996;3(2):S32
[25]张军民,王连递,高振川等.日粮中添加谷氨酰胺对早期断奶仔猪抗氧化能力的影响[J].畜牧兽医学报,2002,33(2):105-109
[26]Dritz,S.S.,K.Q.O wen and R.D.Goodband.1996.Influence of ipopolysaccharideinduced immune challenge and diet c omplexity on growth performance and acutephase protein production in segregated early-weaned pigs.J.Anim.Sci.74:1620-1628.
[27]Klasing K C,Barnes D M.Decreased amino acid requirements of growing Chicks due to immunologic stress.1988,Nutr,118:1158-1164
[28]Thiel-Cooper R L,Parrish F C Jr,Sparks J C,et al.Conjugated linoleic acid changes swine performance andcarcass composition[J].J Anim Sci,2001,79:18211828
[29]Dugan M E R,Aalhus J L,Jeremiah L E.et al.The effects of feeding conjugated linoleic acid on subsequent pork quality[J].Can J Anim Sci,1999,79:45-51
[30]Klasing K C,Laurin D E,Peng R K,et al.Immunologically mediated growth depression in chicks:Influence of feed intake,corticosterone and interleukin-1[J].J Nutr,1987,117:1629-1637
[31]Webel D M,Finck B N,et al.Time course of increased plasma cytokines cortisol and urean itrogen in pigs following intraperitoneal injection of lipopoly saccharide.1997,nim.Sci.75:1514-1520
[32]赖长华,尹靖东,李德发,等.共轭亚油酸对免疫应激仔猪生长抑制的缓解作用.中国畜牧杂志,2005,41(2):6-9
[33]宋林生,邱丽华,蔡中华,等.花鲈肿瘤坏死因子基因及其在LPS和病毒刺激下表达的研究.High TechnOlogy Letters.2004,14(11):81-86
[34]Besedovsky,H,D.R.Adriana,S.Ernst and C.A.Dinarello.1986.nununoregulatory feedback between in terleukin-1 and glucocorticoid hormones.J.Science.233:652-654.
[35]Carroll,J.A.,K.J.Touchete,R.L.Matteri,C.J.Dyer,and G.L.Allee.2002.Efect of spraydried plasma and lipopolySaccharide exposure on weaned pigs:Ⅱ.Efectson the hypothalamicpituitaryadrenal axis of weaned pigs.J.Anim.Sci.80:502-509.
[36]Parrot,R.F.,S.V.Vellucci,and J.A.Goode.1997.Interrelated adrenocortical and neurohypo-Phy sical responses Associated with fever in endotoxin-treated pigs.Am.J.Physiol.273:R1046-R1052.
[37]Ruane N M,Nolan D T,Rotllant J,et al.Experimental exposure of rainbow trout Oneorhynch sykiss(Walbaum) to the infective stages of the sea louse Lepeophtheirus saimonis(Kroyer)influences the physiological response to an acute stressor.Fish & Shell fish Immunology,2000,10:451-463.
[38]Fong,Y,L.L.Moldawer and F.Lowry.1989.Cachectin/TNF or IL-1 induces cachexia with redistribution of body Proteins.Am.J.Physiol.R659-R665.
[39]李建涛,蔡辉益,刘国华等.免疫应激对肉仔鸡采食量、能量及氮消化率和内源氮损失的影响.河北农业大学学报,2005,28(6):90-94
[40]Klasing,K C,and B.J.Johnstone.1991.Monokines in growth and development.Poutr.Sci.70:1781-1789.
[41]Jepson M.M,J.M.Pell,P.C.Bates,and D.J.Millward.1986.The efects of endotoxaemia on protein metabolism in skeletal muscle and liver of fed and fasted rats.J.Bi-ochem.235:329-336.
[42]Memon R A,Feingold K R,Grunfeld C.The efect of cytokines on intermediary Metabolism [J].Endocrinologist,1994,4:59-63
[43]Wright,K.J.,R.Balaji,C.M.HiII,S.s.Dritz,E.L.Knoppel,audJ.E.Minton.Integrated adrenal,soma-Tot-ropic,and immune responses of g owing pigs to treatment with lipopolysaccharide.J.Amim.Sci.,2000,78:1892-1899
[44]杨汉春主编.动物免疫学口[M].北京:中国农业大学出版社,2003
[45]Balaji,R.,K.J.Wright,C.M.Hil],S.S.Dritz,E.L.Knoppel,and J.E.Minton.Acute phase responses pigs challenged orally with Salmonella typhimurium.J.Anim.Sci.,2000,78:1885-1891
[46]白满喜,蒋朱明.谷氨酰胺双肽对大鼠肠切除后细菌移位的影响.中华医学杂志,1996,76(2):116-119
[47]龚时文,张明.谷氨酰胺对大鼠急性坏死性胰腺炎肠道细菌移位的影响.中国急救医学,2000,20(11):640-641
[48]刘涛,周诗其,程学慧等.外源性谷氨酰胺和谷氨酸对早期断奶仔猪肠粘膜形态、结构和小肠吸收功能及骨骼肌中DNA、RNA浓度的影响.中国兽医学报,2003,2391):62-65
[49]石慧琳,程义勇,李树田等.补充谷氨酰胺对束缚大鼠肠屏障功能的影响.营养学报,2004,26(1):31-35
[50]范志勇,王康宁,周定刚.EGF、Gln和pGRF基因质粒对早期断奶仔猪肠道发育的影响.畜牧兽医学报,2006,37(5):457-463
[51]席鹏彬,林映才,蒋宗勇等.谷氨酰胺二肽对断奶仔猪生长、免疫、抗氧化力和小肠粘膜形态的影响.动物营养学报,2007,19(2):135-141
[52]ButIer,EJ.,M.J.,Curtis,and E.G:,Harry.Escherichia coli endotoxin as a stressor in the domestic fowl.Research in veterninary Sci.,1977,23:20-23
[53]Butts,C.A.,Moughan,P.J.,Smith,W.C.Endogenous amino acid flow at the terminal ileum of the rat under conditions of peptide atimentation.J.Sci.Food Agri.,1991,55:175-187
[54]Butts,C.A.,Moughan,P.J,Smith,W.C.,Reynolds,GW.,and Garrick,D.J.The efects of food,dry matter intake on endogenous ileal amino acid excretion determined under peptide alimentation in the 50kg liveweight pig.J.Sci.Food Agri.,1993,62:235-243
[55]Chung,T.K.,and D.H.Baker.Apparent and true amino acid digestibility of a crystalline amino acid mixture and of casein:comparison of values obtained with ileal-cannulated pigs and cecectomized cockerels.J.Anim.Sci.,1992,70:3781-3790.
[56]Chung,T.K,J.Khajareen,and D.R.Zimmerman.Efect of dietary threonine on growth performance and carcass Characteristic of broiler chickens raised at hight Emperatures.In:proceedings of the World's Poultry Congress,New Delhi,India,1996,p.145
[57]Coats,M.M.,FuIIer,r.,et al.A comparision of the growth of chickens in the gustafsson germfree paratus and in conventional environment,with and w ithout dietary supplements of penicillin.Br.J,Nutr.,1963,17:141-150
[58]Colley,C.M.,Fleck,A,Googe,A.W,Muller,B.R.,and Myers,M.A.Early time course of acute phase protein response in man,J.Clin.Path.,1983,36:203-207
[59]周吕.胃肠生理学-基础与临床(M].北京:科学出版社,1991,98-105
[60]Yagoob P,Calder PC.Glutamine requirement of proliferatin T-lymphocytes[J].Nutrition,1997,13:646-651
[61]黄耀凌,邹思湘,陈伟华.谷氨酰胺对小鼠免疫功能的影响[J].营养学报,2001,23(4):363-364
[62]杨俊涛.谷氨酰胺与免疫细胞[J].国外医学创伤与基本问题分册,1997;18:6
[63]林燕.谷氨酰胺对幼建鲤肠道功能和免疫力的影响.四川农业大学:硕士学位论文2005
[64]舒晓亮,曹翔,耿珊珊等.谷氨酰胺对严重创伤手术后患者应激反应和免疫功能的影响.同济大学学报:医学版,2005,26(6):32-34
[65]徐文萍,嵇月娥,施文艳,曹永安.补充谷氨酰胺对急性应激大鼠脑组织GSH、SOD和MDA 含量的影响.中国应用生理学杂志,2003,19(4):415-416
[66]Goldstein L,Schrock H,Cha CJ.Relationship of muscole glutamine to renal ammonia metabolism[J].Biochem Soc Trans,1980,8:509-510
[67]戴四发,金光明,王立克,柳卫国.Gln对肉用仔鸡免疫器官生长发育和成活率的影响.中国畜牧杂志.2003,039(004):25-27
[68]戴四发,王立克等.外源Gln对AA肉鸡肠道早期生长发育的影响.中国饲料.2002.000(016):12-13
[69]Yu B,You Z,Wang S.Enhancement of gut immune function by early enteral feeding enriched with L-glutamine in severe burned miniswines.Zhonghua Zheng Xing Shao shang wai Ke Za Zhi.1996,Mar;12(2):98-100.
[70]刘伟敏,刘忠臣,李新艳,唐光武.Gln肽在断奶仔猪饲料中的应用.饲料博览,2003,000(006):29-30
[71]谈定玉.大黄及谷氨酰胺对肠缺血再灌注大鼠肠道保护作用及其机理研究.[学位论文].南京中医药大学,2006
[72]王维.丙氨谷氨酰胺对大鼠急性肺损伤的影响及其机制.[学位论文].广西医科药大学,2006
[73]Balm,P.H.M,.Stress Physiologyin Animals.Sheffietd Academic Press,Sheffield,England.999.
[74]李静.猪热应激反应适宜监测指标初探[D].沈阳农业大学,2003.
[75]杨春花,黄烽.高敏C反应蛋白研究进展[J],中华风湿病学杂志,2004,8(12):755-758.
[76]计红,杨焕民,吴永魁.热应激蛋白研究进展及其应用前景[J],生物学杂志,2005,22(4):1-3.
[77]张根军.生长猪热应激反应模型及适宜监测指标筛选[D].中国农业科学研究院,2004.
[78]杨钢,席正雄,万瑜等.应激时大鼠血浆、脑、心血管及肾上腺血管紧张素Ⅱ含量的变化[J],生理学报,1993,45(5):505-509.
[79]Mahapatra MS,Mahata SK,Maiti B.Effect of stress on serotonin,norepinephrine,epinephrine and corticosterone contents in the soft-shelled turtle.Clin Exp Pharmacol Physiol.1991,18(10):719-724.
[80]Ray PP,Chaudhuri-Sengupta S,Maiti BR.Adrenocortical involvement during diverse stress in soft-Shelled turtle(Lissemys p.punctata) Bonnoterre.Indian J Exp Biol.2004,42(6):620-625.
[81]Rudy M.Ortiz:Rhonda M.Pattersonb;Charles E.Wadea;F.Michael Byersc.Effects of acute fresh water Exposure on water flux rates and osmotic responses in Kemp's ridley sea turtles (Lepidochelys.kempi) Comparative Biochemistry and Physiology.Part A:Molecular &Integrative Physiology.2000,127,(1):81-87
[82]Ray PP,Maiti BR.Adrenomedullary Hormonal and Glycemic Responses to High Ambient Temperature in the Soft-Shelled Turtle(Lissemys punctata punctata).General and Comparative Endocrinology.2001,122(1):17-22.
[83]Ray PP,Maiti BR.Interrenal responses to high ambient temperature in softshelled turtle,(Lis-semys punctata Punctata).Indian J Exp Biol.2003,41(8):880-884.
[84]Jessop,T.S.,Hamann,M.,Read,M.A.,Limpus,C.J.Evidence for a hormonal tactic maximining green Turtle ReproDuction in response to a pervasive ecological stressor.General and Comparative Endocrinology.2000,118:407-417.
[85]Gregory,L.F.,Gross,T.S.,Bolten,A.B.,Bjorndal,K.A.,Guillette,L.J.Plasma corticosterone concentrations associated with acute captivity stress in wild logger head sea turtles(Caretta caretta).General and Comparative endocrinology.1996,104:312-320.
[86]Gregory L.F.,Schmid,J.R.Stress Responses and Sexing of Wild Kemp's Ridley Sea Turtles (Lepidochelys Kempii) in the Northeastern Gulf of Mexico.General and Comparative Endocrinology.2001,124(1):66-74.
[87]William B.Cash.,Rebecca L.Holberton.,Scott S.Knight.Corticosterone Secretion in Response to Capture and Handling in Free-Living Red-Eared Slider Turtles.General and Comparative Endocrinology.1997,108(3)427-433.
[88]Ott,J.A,Mendonc,M.T.,Guyer,C,,Michener,W,K.Seasonal changes in sex and adrenal teroid hormones of gopher tortoises(Gopherus polyphemus).General and Comparative Endocrinology.2000,117(2):299-31.
[89]Zhou Xianqing,Xie Mengxia,Niu Cuijuan,Sun Ruyong.The effects of dietary vitamin C on growth,liver Vitamin C and serum cortisol in stressed and unstressed juvenile soft-shelled turtles (Pelodiscussinensis).Comparative Bioch-emistry and Physiology Part A:Molecular &Integrative Physiology.2003,135(2):263-270.
[90]Zhou,Xianqing.,Niu,Cuijuan.,Sun,Ruyong.The effects of vitamin E on antiacid stress ability in juvenile soft-shelled turtles(Pelodiscus sinensis).Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology.2004,137(4):299-305.
[91]Jessop,T.S.Modulation of the adrenocortical stress response in marine turtles(Cheloniidae):evidence for a hormone tactic maximizing reproductive investment.J.Zool.Lond.2001,254:57-65.
[92]Valverde,R.A.,Owens,D.W.,Mackenzie,D.s.,Amoss,M.s.Basal and stress-induced corticosterone in olive Ridley sea turtles(Lepidochelys olivacea) in relation to their mass nesting behavior.J.Exp.Zool.1999a,284:652-662.
[93]Valverde,R.A.,Owens,D.W.,Mackenzie,D.s.,Amoss,M.s.Adrenal responsiveness during of the clive ridley sea turtle(Lepidochelys olivacea).Am.Zool.1999b,32A,21.
[94]Jessop,T.s.,Knapp,R,Limpus,C.J.,Whittier,J.M.Dynamic endocrine responses to stress:evidence for energetic constraints and status dependence of breeding in male green turtles.Gen.Comp.Endocrinol.2002,126:59-67.
[95]Aguirre,A.A.,Balazs,G.H.,Spraker,T.R..Adrenal and hematological responses to stress in juve-nile Greenturtles(Chelonia mydas) with and without fibropappillomas.Physiol Zool.1995,68:831-854.
[96]Beatrix G Schramm.,M Casares.,Valentine A.Lance.Steroid Levels and Reproductive Cycle of the Galapagos Tortoise(Geochelone nigra) Living under Seminatural Conditions on Santa Cruz Island(Galapagos).General and Comparative Endocrinology.1999,114(1):108-120.
[97]周显青,牛翠娟,孙儒泳.黄芪和酸应激对中华鳖幼鳖血清补体C3和C4含量的影响[J],动物学研究,2002,23(2):177-180.
[98]周显青,牛翠娟,孙儒泳.饵料维生素E含量对酸应激中华鳖幼鳖血清补体C_3和C_4的影响[J],动物学研究,2003a24(6):452-456
[99]周显青,牛翠娟,孙儒泳.维生素C和酸应激对中华鳖幼鳖血清补体C_3和C_4含量的影响[J],动物学报,2003b,49(6):769-774.
[100]周显青,谢孟峡,牛翠娟,孙儒泳.维生素C和E合用对应激和非应激中华鳖幼鳖生长、肝脏维生素C和E以及血清皮质醇含量的影响[J],动物学报,2004a,50(2):158-164.
[101]陈欣然,牛翠娟,蒲丽君.慢性氨氮暴露对中华鳖生长及非特异性免疫功能韵影响[J],北京师范 大学学报:自然科学版,2006,42(3):300-304.
[102]Willmore WG,Storey KB.Multicatalytic proteinase activity in turtle liver:responses to anoxia stress and recOvery Biochem Mol Biol Int.1996,38(3):445-51.
[103]Willmore WG,Storey KB.Purification and properties of the glutathione S-transferases from the anoxiatolerant turtle(Trachemys scripta elegans).FEBS J.2005,272(14):3602-3614.
[104]Hossein Mehrani;Kenneth B.Storey.Enzymatic control of glycogenolysis during anoxic submergence in the Fresh water turtle(Trachemys scripta).The International Journal of Biochemistry & Cell Biology.1995,27(8):821-830.
[105]Bagatto,B,Henry,RP.Exercise and forced submergence in the pond slider(Trachemys scripta)and softshell turtle(Apalone ferox):influence on bimodal gas exchange,diving behaviour and blood acid-base status.Journal of Experimental Biology.1999,202(3):267-278.
[106]Jackson DC,Warburton SJ,Meinertz EA,Lawler RG,Wasser JS.The effect of prolonged anoxia at 3 degrees C on tissue high energy phosphates and phosphodiesters in turtles:a 31P-NMR study.J Comp Physiol(B);1995,165(1):77-84.
[107]Wasser JS,Heisler N.Effects of anoxia on intracellular free Ca~(2+) in isolated cardiomyocytcs from turtles.Comp Biochem Physiol A Physiol.1997,116(4):305-12.
[108]Bobb VT,Jackson DC.Effect of graded hypoxic and acidotic stress on contractile force of heart muscle from ypoxiatolerant and hypoxia-intolerant turtles.J Exp Zoolog A Comp Exp Biol.2005,303(5):345-53.
[109]Attilio Arillo;Emilio Balletto;Maria A.Cherchi.Quantitative variations of some compounds of the T.C.A.-cycle in specimens of Testudo hermanni grnelin after cxposure to hyperthermic conditions.Journal of Thermal Biology 1997,2(3):131-133.
[110]Michel Cahanac,Cecile Bernieri..Behavioral rise in body temperature and achycardia by handling of a Turtle(Clemmys insculpta).Behavioural Processes.2000,49(2):61-68.
[111]Rotermund A.J;Privitera C A..The effects of induced cold torpor on ATPase activity of the turtle,Pseudemys(chrysemys) picta.Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology.1972,41(3):511-520.
[112]Emilio Balletto,Maria A.Cherchi,Federico Melodia,Paola Mensi.Temperaturcdependent thyroid hormone Bios-ynthesis in Testudo horsfieldi Gray.General and Comparative Endocrinology.1979,39(4):548-551.
[113]Baccari GC,Minucci S,Di Matteo L.The orbital glands of the terrapin Pseudemys scripta in response to osmotic stress:a light and electron microscope study.J Anat.1993,183(1):21-33.
[114]周显青,牛翠娟,李庆芬等.光照强度对中华鳖稚鳖摄食和生长的影响[J],动物学报,1998a,44(2):157-161.
[115]周显青,牛翠娟,李庆芬等.光照度对中华鳖稚鳖能量转换的影响[J],北京师范大学学报(自然科学版),1998b,34(2):248-251.
[116]周显青,牛翠娟,李庆芬.光周期对中华鳖稚鳖摄食,生长和能量转换的影响[J],生态学报,1999a(3):383-387.
[117]Guillette Jr.,L.J.,Cree,A.,Rooney,A.A.biology of stress:interactions with reproduction,immunology and Inter mediary etabolism,in:Warwick,C.,Frye,F.L,Murphy,J.B.(eds),Health and welfare of Captive Reptiles,Chapman and hall,London,pp.1995,32-81.
[118]Mahmoud,I.Y;Licht,Paul.Seasonal Changes in Gonadal Activity and the effects of Stress on Reproductive Hormones in the Common Snapping Turtle(Chelydra serpentina).General and Comparative Endocrinology.1997,107(3):359-372
[119]Licht P,Breitenbaeh GL,Congdon JD.5Seasonal cycles in testicular activity,gonadotropin,and thyroxine in the painted turtle(Chrysemys picta) under natural conditions.Gen Comp Endocrinol.1985,59(1):130-9.
[120]Huot-Daubremont C.;Bradshaw S.D.;Bradshaw F.J.;Kuchling G;Grenot C.J.variation of plasma sex steroid concentrations in wild and captive populations of hermann's tortoise(Testudo hermanni hermanni) in Southern France.General and Comparative Endocrinology.2003,130(3):299-307.
[121]Jessop,T.S.,Limpus,C.J.,Whittier,J.M.Plasma steroid interactions during highdensity green turtle nesting and associated disturbance.Gen.Comp.Endocrinol.1999,115,90-100.
[122]Prassack SL,Bagatto B,Henry RP.Effects of temperature and aquatic P(O_2) on the physiology and behaviour of Apalone ferox and Chrysemys picta.The Journal of Experimental Biology.2001,204,2185-2195.
[123]Jon P Costanzo;Jaequeline D Litzgus;Richard E LeeJr.Behavioral responses of hatchling painted turtles(ChryseMys picta) and snapping turtles(Chelydra serpentina) at subzero temperatures.Journal of Thermal Biology.1999,24(3):161-166.
[124]周显青,牛翠娟,李庆芬.光照对水生动物行为的影响[J],动物学杂,1999b,34(2):45-48.
[125]Richard W.Brill;George H.Balazs;Kim N.Holland;Randolph K.C.Chang;Scott Sullivan;John C.George.Daily movements,habitat use,and submergence intervals of normal and.tumor-bearing juvenile green turtles(Chelonia mydas L).within a foraging area in the Hawaiian islands.Journal of Experimental Marine Biology and Ecology.1995,185(2):203-218.
[126]王军,赵景宏,粟永萍.糖皮质激素受体调控与创伤应激紊乱关系的研究进展[J],中华创伤杂志,2001,17(8):508-510.
[127]Rice M.E;Forman R.Chen E;B.T;Avshalumov M.V;Cragg S.J;Drew K.L.Brain antio-xidant regulation in mammals and anoxia-tolerant reptiles:balanced for neuroprotection and neuromodulation.Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology.2002,133(4):515-525.
[128]周显青,牛翠娟,孙儒泳.维生素E对中华鳖幼鳖生长、肝脏维生素E以及血清皮质醇含量的影响[J],动物学报,2003c,49(1):40-44.
[129]周显青,牛翠娟,孙儒泳.维生素C和E合用对中华鳖幼鳖非特异性免疫功能的影响(J),水生生物学报,2004b,28(4):356-360.
[130]周显青,牛翠娟,孙儒泳.黄芪对中华鳖免疫和抗酸应激能力的影响[J],水生生物学报,2003d,27(1):110-112.
[131]肖明松,王志耕,崔峰,等.果寡糖和糖萜素对中华鳖生长性能及免疫功能的影响.水利渔业,2004,24(3):20-22
[132]刘宗英,姚鹃,陈昌福,等.高活性干酵母对中华鳖非特异性免疫功能和抗病力的影响.华中农业大学学报,2005,24(2):192-196
[133]汪成竹,姚鹃,吴凡,等.免疫多糖(酵母细胞壁)对中华鳖非特异性免疫功能的影响.华中农业大学学报,2006,25(4):421-425
[134]覃川杰,汪成竹,陈晓辉等.茯苓多糖对中华鳖非特异性免疫功能的免疫调节作用.淡水渔业,2006,36(6):40-43
[66]王亭亭,蔡完其.饲料中添加花粉和酵母硒对中华鳖幼鳖生长和非特异性免疫功能的影响.上海水产大学学报,2005,14(2):97-102
[135]宋超,牛翠娟,陈欣然.壳聚糖对急性氨氮胁迫下中华鳖稚鳖非特异性免疫反应的影响.动物学报,2007,53(2)270-277
[136]王亭亭,蔡完其.饲料中添加花粉和酵母硒对中华鳖幼鳖生长和非特异性免疫功能的影响.上海水产大学学报,2005,14(2):97-102
[137]刘至治,蔡完其,季高华,等.几种免疫增强剂对中华鳖红细胞数量及免疫功能的影响.上海水产大学学报,2006,15(1):1-15
[138]赵燕,代兵,李传普等.半胱胺对中华鳖生长性能和非特异性免疫功能的影响研究.动物营养学报,2007,19(3)305-310
[139]陈昌福.柱状嗜纤维菌脂多糖对鲤不同组织中离体培养淋巴细胞转化率的影响.华中农业大学学报,1998,17(1):66-70
[140]陈昌福,吴志新.三种爱得华氏菌脂多糖对日本鳗鲡免疫原性的比较.水生生物学报,1998,22,增刊:126-131
[141]陈昌福,楠田理一.三种鳜对柱状嗜纤维菌脂多糖免疫应答的比较研究.华中.农业大学学报,1999,18(3):252-255
[142]陈昌福,曾妍雄,楠田理一.接种不同种类病原菌脂多糖对翘嘴鳜细菌性败血症的免疫保护力比较.华中农业大学学报,2000,19(4):377-380
[143]陈昌福,罗宇良,曾妍雄.不同血清型柱状嗜纤维菌脂多糖蛋白质成分及其对鳜免疫原性影响的研究.华中农业大学学报,2000,19(5):483-486
[144]曾慷,何建国,王晓红,翁少虹,左涛.传染性脾肾坏死病毒,Con A和LPS对鳜的头肾淋巴细胞转化的影响.水产学报,2001,25(5):464-468
[145]黄辨非,陈超然,毛之娟,陈昌福.同腹异育银卿对鱼类3种致病弧菌脂多糖的免疫应答.华中农业大学学报,2002,21(1):59-63
[146]陈昌福,陈超然.鱼类三种致病菌的粗脂多糖对异育银卿的免疫原性.水生生物学报,2002, 26(5):483-488
[147]Trapnell,JE.Pathophysiology of acute pancreatitis.World J Surg,1981,5(3):319-327
[148]田文霞,徐彤,郑明学等.实验性免疫应激雏鸡免疫器官抗体生成细胞的动态变化研究.山西农业大学学报,2000,(3):232-235
[149]徐彤,高文伟,任家琰等.实验性免疫应激对雏鸡血液激素与免疫功能的影响.华北农学报,2002,17(1):114-118
[150]Johnson R W.Inhibition of growth by proinflammatory cytokines:An intergrated view.Journal of Animal Science,1997,75:1244-1255
[151]林清华.免疫学实验[M].武汉:武汉大学出版社,1995:188-195
[152]Knodel M,Kuss AW,Berberich,et al.Blimp-1 over-expression abrogates IL-4 and CD40 mediated of terminal B cell differentiation but arrests isotype switching[J].Eur J Immunol,2001,31:1972-1980
[153]简纪常,吴婷婷,杨弘,董在杰,曹莹.中华鳖体液免疫应答的初步研究.中国水产,1998,12(4):6-10
[154]简纪常.中华鳖血清中抗体含量的研究.动物学研究,2001,22(2):163-165
[155]Sanders M M,Ken C.Glutamine is a powerful efector of heat shock protein expression in Drosophila Kccens[J].J Cell Physiol,1992,146:180.
[156]W.ischaneyer P E,Kahana M D,Wolfson R,et al.Glutamine induces heat shock protein and protects against endotoxin shock in the rat[J].J Appl Physiol,2001,90,2403.
[157]张丽英.饲料分析及饲料质量检测技术(第二版)[M].北京:中国农业大学出版社,2003:56:59
[158]陈静,刘显军,边连全,石娇.谷氨酰胺对早期断奶仔猪生长性能和免疫性能的影响.西北农业学报,2006,15(4):58-62
[159]高春生,王艳玲,杨国宇,等.谷氨酰胺(Gln)对早期断奶仔猪生长性能和腹泻的影响.中国农学通报.2006年22(4):12-14
[160]范志勇,王康宁,周定刚.EGF、Gln和pGRF基因质粒对早期断奶仔猪生产性能、免疫功能及相关激素水平的影响.畜牧兽医学报,2005,36(9):893-899
[161]果磊,贺光照,黄崇本.谷氨酰胺对烧伤大鼠肠道粘膜损伤的保护作用.重庆医科大学学报,2002,27(4):403-406.
[162]陈秋生,聂其灼.中华鳖脾脏白髓的组织结构与组织化学特性.甘肃农业大学学报,1993,28(3):220-223
[163]陈秋生,聂其灼.中华鳖脾脏的显微与亚显微结构研究.南京农业大学学报,1995,18(4):91-97
[164]赵万鹏,别立洁.鳖脾脏的正常组织学和病理组织学.信阳师范学院学报(自然科学版),2003,16(1):45-48